Method for inhibiting extracellular antibiotic resistance gene transformation in water by using biochar
A technology of antibiotic resistance and gene transformation, applied in the new application field of biochar in inhibiting the transformation of antibiotic resistance genes in water bodies, can solve the problem of accelerating the generation and spread of antibiotic-resistant bacteria and antibiotic resistance genes, and reducing the risk of antibiotic treatment for humans. and the ability of animal pathogens to infect, etc., to achieve the effect of huge specific surface area and rich pore structure
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[0036] In the preparation process of the biochar, the pyrolysis temperature of the biochar is 300°C-700°C, and the pyrolysis time is 5-7h. The pyrolysis temperature of the further biochar is preferably 300°C, and the pyrolysis time is preferably 6h. The biomass used for pyrolysis can be rice straw or peanut shell, preferably rice straw.
[0037] After the biochar is obtained, the preparation method of making it into a biochar leaching solution is as follows: mixing the biochar with a pH buffer solution, shaking fully, passing the solution through a 0.45 μm filter membrane to obtain a biochar leaching solution. The pH buffer used is TB buffer, the shaking temperature is room temperature, the shaking speed is 150 r / min, and the shaking time is 3 h. The role of TB buffer is to maintain a constant pH of the extract solution to facilitate the growth of microorganisms.
[0038] Through the above method, biochar can be used to inhibit the transformation of extracellular antibiotic r...
Embodiment 1
[0041] Embodiment 1: the preparation of biochar
[0042] Present embodiment 1 mainly realizes the preparation of biochar, and its preparation process is as follows:
[0043] Two kinds of raw materials were selected for biochar preparation, rice straw (Rice) and peanut shell (PeanutS). Before preparation, the three kinds of biomass were washed with water and dried in an oven at 80°C for 48 hours; after drying, the biomass was taken out, pulverized in a pulverizer, passed through a 50-mesh sieve, and stored in a sealed bag for dry storage. During preparation, the crushed and sieved biomass powder is put into a 250ml ceramic crucible, compacted and covered with a lid, and placed in a muffle furnace for high-temperature pyrolysis. The heating rate of the muffle furnace is 5°C / min, two target temperatures are set, 300°C and 700°C, and the residence time is 6h. After the pyrolysis in the muffle furnace is completed and cooled naturally to room temperature, take out the crucible, gri...
Embodiment 2
[0044] Embodiment 2: Preparation of resistant plasmid and culture medium
[0045] In this example, the resistance plasmid and culture medium used in subsequent examples are mainly prepared, and the preparation process is as follows:
[0046] The pUC18 plasmid carrying the ampicillin antibiotic resistance gene was extracted from Escherichia coli (E. Coil), and the kit used was the high-purity plasmid extraction kit (DP116) produced by Tiangen Company. The concentration and purity of the proposed plasmid solution were detected by NanodropOne, A260 / A280>1.8, indicating that the purity was relatively pure, and the concentration of the plasmid solution was 145mg / L. In this example, Escherichia coli (E. Coil) was used as the recipient cell, LB medium was used as the growth medium of E. Coil, and competent cells (E. Coil DH5α) were purchased from Shanghai Sangong.
[0047] LB liquid medium formula: tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L;
[0048] LB solid medium formula: tr...
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